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E86. In the methods of Embodiments 81 to 85, the improvement further comprising measuring reduction of in vitro expression of a target protein of the molecule. The present invention relates to methods for selecting therapeutic molecules that have reduced toxic side effects. The present disclosure provides a method of testing or determining in vivo acute neurotoxicity of a molecule comprising measuring oscillations in intracellular free calcium concentration ("calcium oscillations") in vitro in neuronal cells which are in contact with the molecule. One central concern when administering therapeutic molecules to a subject is the potential neurotoxicity of the molecules. The ASO and other oligomerss that behaved similarly, were shown to negatively impact central nervous system (CNS) network activity in vivo and electrophysiologic spontaneous neuronal activity in vitro (data not shown). FIG. 4: shows the impact of Tau antisense oligonucleotide on spontaneous calcium oscillations in primary neurons. E2. The method of embodiment 1, wherein the calcium oscillations of the molecule are compared to the calcium oscillations in neuronal cells that are not exposed to the molecule ("control cells"). E43. The method of embodiment 42, wherein the behavioral test score is measured by administering the molecule to a mammal and grading the mammal's` behavioral performance.

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